The study of ex vivo phagocytosis via flow cytometry requires that one distinguish experimentally between uptake and adsorption of fluorescently labeled targets by phagocytes. Removal of the latter quantity from the analysis is the most common means of analyzing such data. Because the probability of phagocytosis is a function of the probability of adsorption, and because partially quenched fluorescence after uptake often overlaps with that of negative controls, this approach is suboptimal at best. Here, we describe a numerical analysis model which overcomes these limitations. We posit that the random adsorption of targets to macrophages, and subsequent phagocytosis, is a function of three parameters: the ratio of targets to macrophages (m), the mean fluorescence intensity imparted to the phagocyte by the internalized target (alpha), and the probability of phagocytosis per adsorbed target (p). The potential values of these parameters define a parameter space and their values at any point in parameter space can be used to predict the fraction of adsorption(+) and [adsorption(-), phagocytosis(+)] cells that might be observed experimentally. By systematically evaluating the points in parameter space for the latter two values and comparing them to experimental data, the model arrives at sets of parameter values that optimally predict such data. Using activated THP-1 cells as macrophages and platelets as targets, we validate the model by demonstrating that it can distinguish between the effects of experimental changes in m, alpha, and p. Finally, we use the model to demonstrate that platelets from a congenitally thrombocytopenic WAS patient show an increased probability of ex vivo phagocytosis. This finding correlates with other evidence that rapid in vivo platelet consumption contributes significantly to the thrombocytopenia of WAS. Our numerical analysis method represents a useful and innovative approach to multivariate analysis.

PubMed | Sloan Kettering Cancer Center, Fuller Theological Seminary, University of Memphis, The New School for Social Research and Memphis Veterans Administration Medical CenterType: Comparative Study | Journal: Death studies | Year: 2014

This study examined patterns of making meaning among 155 parents whose children died from a variety of violent and non-violent causes. Findings indicated 53% of violent loss survivors could not make sense of their loss, as compared to 32% of non-violent loss survivors. Overall, there was overlap in sense-making strategies across different causes of death, with many parents invoking spiritual and religious meanings and the cultivation of empathy for the suffering of others. Nonetheless, violent loss survivors described the imperfection of the world and brevity of life more frequently in their narrative responses than parents who lost a child to natural causes, who in turn were more likely to find benefit in the loss in terms of personal growth. Violent loss survivors--and especially those losing a child to homicide--also reported enhanced appreciation of life more frequently than survivors of non-violent losses, and surviving a childs suicide was specifically associated with a change in priorities in the sample. Findings are discussed in terms of common and distinctive themes in meaning making that clinicians may encounter when working with parental bereavement, and the implications these carry for finding spiritual and secular significance in a traumatic loss.

Unlike anemias, most thrombocytopenias cannot be separated into those due to impaired production and those due to accelerated consumption. While rapid clearance of labeled platelets from the bloodstream can be followed in thrombocytopenic individuals, no model exists for quantitatively inferring from autologous or allogeneic platelet consumption data what changes in random consumption, lifespan dependent consumption, and platelet production rate may have caused the thrombocytopenia. Here we describe a numerical analysis model which resolves these issues. The model applies three parameter values (a random consumption rate constant, a lognormally-distributed platelet lifespan, and the standard deviation of the latter) to a matrix comprising a series of platelet cohorts which are sequentially produced and fractionally consumed in a series of time intervals. The cohort platelet counts achieved after equilibration of production and consumption both enumerate the population age distribution and sum to the population platelet count. Continued platelet consumption after production is halted then serves to model in vivo platelet consumption data, with consumption rate in the first such interval defining the equilibrium platelet production rate. We use a least squares fitting procedure to find parameter values which best fit observed platelet consumption data obtained in WT and thrombocytopenic WASP(-) mice. Equilibrium platelet age distributions are then 'grafted' into the matrix to allow modeling of the consumption of WT platelets in WASP(-) recipients, and vice versa. The optimal parameter values obtained indicate that random WT platelet consumption accounts for a larger fraction of platelet turnover than was previously suspected. Platelet WASP deficiency accelerates random consumption, and a trans effect of recipient WASP deficiency contributes to this. Application of the model to clinical data will allow distinctions to be made between thrombocytopenias due primarily to impaired platelet production and those due to acceleration of random or lifespan-dependent platelet consumption.

Attachment theory has become a primary framework for understanding adjustment to traumas. In a convenience sample of 157 U.S. service members from the Afghanistan and Iraq wars seeking health care services at a Veterans Administration (VA) hospital, this study examined (a) the impact of attachment characteristics on several key mental health symptoms in this new generation of veterans, (b) the relative frequencies of prominent attachment styles in the sample, and (c) how these higher order orientations related to study outcomes. First, with demographic and military background factors in the model, attachment-related anxiety and avoidance were each uniquely associated with posttraumatic stress symptoms (PTSS), psychiatric distress, and alcohol misuse, s = .25 to .60. Second, latent class analysis highlighted the underrepresentation of avoidant veterans of a dismissive type in the sample. Third, of the different possible types of attachment insecurities, veterans with a fearful disorganized orientation were also shown to be particularly vulnerable to PTSS and other problems, even when compared to preoccupied-dependent persons. These findings yield further support for the protective benefit of attachment security, while also suggesting the importance of both attachment anxiety and avoidance in the context of postdeployment adjustment.

Crises in personal meaning may be a critical aspect of psychological maladjustment following combat. Using the newly developed Integration of Stressful Life Events Scale (ISLES), this study examined the role of meaning made of a salient stressor following a combat deployment in Iraq or Afghanistan. In a sample of 169 returning service members, findings supported the distinctiveness of meaning made of a stressor (as assessed by the ISLES), factor analytically and as a correlate of several relevant clinical outcomes. In particular, when the model contained potential confounds and psychiatric symptoms, meaning made in the months and years following combat was uniquely associated with the severity of posttraumatic stress symptoms, = -.39, and referrals for mental health care, = -.69, in the sample. The present findings highlight the importance of the subjective meaning made of traumatic life events and also suggest that posttraumatic stress and other psychiatric symptomatology may not account for the full clinical picture among many service men and women.

Thrombocytopenia caused by rapid platelet consumption contributes to the severe thrombocytopenia of Wiskott-Aldrich syndrome (WAS) and to the milder thrombocytopenia seen in murine WAS. We show that rapid clearance of In-labeled murine WASP(-) platelets correlates with enhanced splenic uptake. Using platelets labeled with a pH-sensitive fluorescent marker (pHrodo), we quantify normal platelet uptake by red pulp macrophages (RPMs), and demonstrate its enhancement after in vivo opsonization of platelets. The spleens of WASP(-) mice contain an increased number of RPM, and rapid clearance of WASP(-) platelets in WASP(-) mice in turn generates an increased number of pHrodo(+) splenic RPMs. To separately assess the platelet intrinsic and recipient-dependent functions involved in the clearance and splenic phagocyte uptake of WASP(-) platelets, we performed crossed pHrodo(+) platelet injection studies (wild type [WT] to WASP(-), WASP(-) to WT). We show that an extrinsic effect of recipient WASP deficiency on the clearance of WASP(-) platelets correlates with increased platelet uptake by RPMs. An intrinsic effect of platelet WASP deficiency on platelet clearance does not, however, correlate with increased total uptake by WT or WASP(-) RPMs. In contrast to other published findings, we find no evidence of a baseline or antibody-induced increase in phosphatidyl serine exposure on WASP(-) platelets. Our findings suggest that an increased number of RPMs in WASP(-) mice contributes significantly to the increased platelet consumption rate in WASP(-) mice. This might explain the consistent efficacy of splenectomy in murine and clinical WAS.